CN108969397A - A kind of active peptides facial mask and preparation method thereof - Google Patents
A kind of active peptides facial mask and preparation method thereof Download PDFInfo
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Abstract
The present invention relates to a kind of active peptides facial masks and preparation method thereof, belong to cosmetic technical field.The invention proposes a kind of active peptides facial masks, it has preferable anti-oxidant, moisturizing action and efficacy, sturgeon fish-skin is mainly used as animal sources, have passed through secondary enzymolysis, membrane separating and purifying has obtained active peptides, be applied to face pack in show preferable skin anti-aging effect.
Description
Technical field
The present invention relates to a kind of active peptides facial masks and preparation method thereof, belong to cosmetic technical field.
Background technique
Currently, anti-aging cosmetics have accounted in facial-care nearly 40% market share, with the prosperity of modern society's information
And the raising of consumer's know-how, nowadays, young woman is also added to anti-aging lotion, cream and essence etc. daily
In nursing procedure, anti-aging product has been expanded in wider array of age level and product scope.Anti-aging product concept constantly refreshes,
Main body trend is natural and high-tech.The elastic force of skin and the source of elasticity are the collagens for accounting for skin corium 70%, corium
Other compositions are elastin laminin, hyaluronic acid etc..Fiber sprout cell is responsible for the synthesis and decomposition of these ingredients, makes skin always
Keep young.Cosmetics peptide is added in skin fiber sprout cell culture solution, culture measures the appreciation rate of cell, discovery two days later
After adding cosmetics peptide, fiber sprout cell is proliferated, and there are interdependences with peptide concentration.Anti-oxidation peptide can also make the saturating of skin
Bright matter acid generative capacity activation.
In recent years, researcher is extracted a variety of anti-oxidation peptides, LeeD [1] from dairy products, animal muscle, bone and skin
Deng report, it is free that the octapeptide (941.43Da) isolated from duck skin by-product removes hydroxyl radical free radical, DPPH free radical, alkyl
The IC50 value of base and ultra-oxygen anion free radical is respectively 32.6 μ g/mL, 22.7 μ g/mL, 55.1 μ g/mL and 49.8 μ g/mL, and
Have and alleviates the effect of the hepatocellular injury as caused by alcohol；Anti-oxidation peptide is prepared from frontal bone albumen in Li Jiaojiao [2] etc., surpasses
Oxygen anion free radical clearance rate reaches 64.14%.In recent years, major cosmetics company release one after another polypeptide series products [3,
4].Lancome modeling face Firm series contains polished rice peptide, can consolidate the wave structure of skin articulamentum, accurately pulling facial skin makes
Skin restores elasticity and cohesiveness.SK-II Firm re-surface intensive wrinkle correct cream, which has contained the collagens such as palm tetrapeptide -3 and atelocollagen, to be repaired
The factor is protected, microgroove and wrinkle can be quickly desalinated.Venin Royale is proposed a containing 27 kinds of native peptides, neuropeptide and mind
Antisenility skin care product K arin Herzog Vitamin H Face Cream through toxin.Tau Collagen Mask is
The face cream of a collagen containing high concentration hydrolysis, hyaluronic acid and squalene, the product can improve in 7~10 min
Texture.
But existing animal sources anti-oxidation peptide remains that DNA purity is high, active needs further increase
Demand.
Bibliography:
[1] Lee S J, Kim Y S, Hwang J W, et al. Purification and characterization
of a novel antioxidative peptide from duck skin by-products that protects
liver against oxidative damage[J]. Food Research International, 2012, 49(1):
285-295.
[2] Li Jiaojiao, Li Cheng, Fu Gang wait enzymatic hydrolysis frontal bone albumen to prepare process optimization [J] food work of anti-oxidation peptide
Industry science and technology, 2013,34 (20): 194-198.
[3] in progress [J] of next lucky, He Congfen, the fourth of the twelve Earthly Branches Dong Yin skin aging mechanism and anti-aging cosmetics
State's beauty medical journal, 2009,18 (8): 1208-1212.
[4] Manso M A, Miguel M, Even J, et al. Effect of the long-term intake of
an egg white hydrolysate on the oxidative status and blood lipid profile of
spontaneously hypertensive rats[J]. Food Chemistry, 2008, 109(2):361-7.
Summary of the invention
The invention proposes a kind of active peptides facial masks, have preferable anti-oxidant, moisturizing action and efficacy, mainly
Using sturgeon fish-skin as animal sources, secondary enzymolysis have passed through, membrane separating and purifying has obtained active peptides, be applied to facial mask
Preferable skin anti-aging effect is shown in cosmetics.
The first aspect of the invention:
A kind of active peptides facial mask, include including matrix and the facial mask liquid being applied in matrix, in the facial mask liquid according to
The following component of weight percent meter: sturgeon fishskin polypeptide 2-5%, whitening composition 0.1-0.3%, thickener 2-4%, emulsifier 3-
3.5%, soluble small molecular 4-6%, preservative 0.02-0.04%, essence 0.005-0.01%, deionized water 85-88%.
In one embodiment, the preparation method of the sturgeon fishskin polypeptide includes the following steps:
Step 1, it pre-treatment: after sturgeon skin is crushed, is soaked in NaCl solution and removes non-collagen, then residue is soaked in
By isopropanol, ether, distilled water mixing and solution in, removal fat, filter out solids；
Step 2, it digests for the first time: solids obtained in step 1 is dispersed in water, HCl is added and adjusts pH to 5.5-6.5,
Then it is digested using bromelain, after enzyme deactivation, adjusting pH to neutrality, obtains first time enzymolysis product；
Step 3, first time enzymolysis product large aperture hyperfiltration treatment: is filtered removal impurity with ultrafiltration membrane；
Step 4, nanofiltration membrane is concentrated: will obtain that ZnCl is added in ultrafiltrated permeation liquid in step 32, reuse molecular cut off 800Da
Nanofiltration membrane be filtered concentration；
Step 5, it digests for second: HCl is added in nanofiltration membrane concentrate obtained in step 4 and adjusts pH to 5.5-6.5, is then made
It is digested with bromelain, after enzyme deactivation, adjusting pH to neutrality, obtains second of enzymolysis product；Successively pass through again
It is concentrated under reduced pressure, spray drying, obtains sturgeon fishskin polypeptide.
In one embodiment, the concentration of NaCl solution is 5-15% in step 1；Isopropanol, ether, distilled water weight
Measuring ratio is 2-4:1-3:6-10.
In one embodiment, the concentration of solids in water is 0.1-0.5wt% in step 2, and enzyme concentration is 2000U/
G substrate, hydrolysis temperature are 50-60 DEG C, and enzymolysis time is 60-120min.
In one embodiment, the molecular cut off of ultrafiltration membrane is 200,000-40 ten thousand Da in step 3.
In one embodiment, the Zn in step 4 in ultrafiltrated permeation liquid2+Concentration is in 0.1-0.5mol/L, nanofiltration membrane
Molecular cut off be 500-1000Da.
In one embodiment, enzyme concentration is 4000U/g substrate in step 5, and hydrolysis temperature is 45-55 DEG C, when enzymatic hydrolysis
Between be 50-70min.
In one embodiment, whitening composition is selected from Pomegranate Peel Extract.
In one embodiment, thickener is selected from pectin, Sodium Hyaluronate, guar gum, xanthan gum, acrylate, carboxylic
The mixing of one or more of methylcellulose, cellulose gum or ethyl cellulose.
In one embodiment, emulsifier is selected from hexadecanyl phosphate sylvite, stearoylketene base sodium glutamate, stearoylketene
One of base-sodium cocoyl glutamate is a variety of.
In one embodiment, soluble small molecular is selected from glycerol, 1,3-BDO, one of propylene glycol or more
Kind.
In one embodiment, preservative is methyl p-hydroxybenzoate, ethyl-para-hydroxybenzoate, para hydroxybenzene first
One or more of propyl propionate, benzoic acid and Phenoxyethanol.
In one embodiment, the matrix is non-woven fabrics.
The second aspect of the invention:
The preparation method of active peptides facial mask, includes the following steps:
Step a disperses a part of deionized water of sturgeon fishskin polypeptide, and emulsifier and soluble small molecular is added, is warming up to
It 40-50 DEG C, stirs evenly；
Step b continuously adds thickener and another part deionized water, adds whitening composition, is warming up to 55-60 DEG C, stirring
Uniformly；
Preservative and essence are added in the solution obtained in stepb, stirs evenly, obtains facial mask liquid by step c；
Step d, by facial mask liquid be coated on matrix on to get.
The third aspect of the invention:
A kind of sturgeon fishskin polypeptide is to be digested to obtain by bromelain using sturgeon fish.
The fourth aspect of the invention:
Purposes of the above-mentioned sturgeon fishskin polypeptide in cosmetics.
The fifth aspect of the invention:
Above-mentioned sturgeon fishskin polypeptide is in the purposes being used to prepare in the medicament for removing DPPH free radical.
The sixth aspect of the invention:
Above-mentioned sturgeon fishskin polypeptide is in the purposes being used to prepare in the medicament for removing hydroxyl radical free radical.
Beneficial effect
The present invention has obtained the sturgeon that a kind of oxidation resistance is strong, activity is high by carrying out enzymatic hydrolysis twice to sturgeon fish-skin
Fish polypeptide, preferable skin effect can be shown in facial mask by applying.
In polypeptide production methods of the invention, for the first time enzymatic hydrolysis purpose be sturgeon fish-skin is tentatively digested, wherein
A part of lower albumen of activity is not easily decomposed, and can be removed the albumen of macromolecular and impurity in subsequent ultra-filtration process
It removes, the activity of polypeptide can be improved；Subsequent nanofiltration process is that first time enzymolysis product is concentrated, due to the mistake in enzymatic hydrolysis
Cheng Zhonghui adjusts the pH of enzymolysis liquid by the way that HCl and NaOH is added, and it is not high to result in polypeptide product purity, can retain NaCl in polypeptide
In, therefore, by the way that Zn is added in ultrafiltration permeate2+, on the one hand since nanofiltration membrane is charged membrane, in the two sides of film, there is electricity
Lotus balances Donnal effect, and because of nanofiltration membrane rejection with higher for divalent ion, Zn2+It can be with polypeptide one
The retention side for residing in nanofiltration membrane is acted, and nanofiltration membrane can force more Na to keep charge balance+Through nanofiltration membrane to infiltration
Saturating side, so that the NaCl content in polypeptide is reduced；On the other hand, since bromelain is to Zn2+More sensitive,
Zn2+It resides in nanofiltration retentate fluid, can further promote the enzymatic hydrolysis of fishskin polypeptide during subsequent secondary enzymolysis, obtain
Polypeptide product activity it is higher；Still further, due to Zn2+There are obvious help, Zn for the bacteriostatic activity of skin surface2+
It resides in and extracts in product polypeptide, mask product can be enable to promote the biocidal property of skin surface.Therefore, in above-mentioned step,
It is achieved that primary enzymolysis and ultrafiltration preliminary screening high activity polypeptide, Zn2+NaCl is promoted to penetrate nanofiltration membrane, Zn2+Reside in nanofiltration
Promote secondary enzymolysis, Zn in concentrate2+Promote facial mask to the overall coordination effect of skin biocidal property.
Detailed description of the invention
Fig. 1 is the ultraviolet spectrogram of sturgeon fishskin polypeptide；
Fig. 2 is the liquid phase exclusion chromatography figure of polypeptide obtained in embodiment 1；
Fig. 3 is the liquid phase exclusion chromatography figure of polypeptide obtained in reference examples 1；
Fig. 4 is the liquid phase exclusion chromatography figure of polypeptide obtained in reference examples 2；
Fig. 5 is comparison of the polypeptide to DPPH free radical scavenging activity；
Fig. 6 is comparison of the polypeptide to hydroxyl radical free radical clearance rate；
Specific embodiment
The preparation of 1 active peptides of embodiment
Pre-treatment: after sturgeon skin is cut, by scraped cleans such as the subcutaneous fat of fish, connective tissues, then being cut into powder, leaching
It steeps and removes soluble non-collagen in 10wt%NaCl solution, filter out residue and be soaked in isopropanol, ether, distilled water again
According in weight ratio 3:2:8 mixed liquor, fat is removed, is dried after filtering, obtains substrate.
Digest for the first time: substrate be configured to the suspension of 0.2wt% in deionized water, then with dilute HCl adjusting pH to
6.0 or so, bromelain is added, enzyme concentration is 2000U/g substrate, digests 90min under the conditions of 55 DEG C, is then gone out for 100 DEG C
Enzyme 10min adds NaOH and adjusts pH to neutrality, obtains first time enzymolysis product；
Large aperture hyperfiltration treatment: molecular cut off is used to be filtered for the ultrafiltration membrane of 300,000 Da first time enzymolysis product, enzyme
It solves product and penetrates ultrafiltration membrane, the lower albumen of activity and big molecular impurity are retained by ultrafiltration membrane, collect penetrating fluid；
Nanofiltration membrane concentration: ZnCl will be added in ultrafiltration membrane penetrating fluid2, make Zn2+Concentration is divided in 0.1mol/L, then using retention
The nanofiltration membrane of son amount 800Da is filtered concentration, and the polysaccharide, NaCl in enzymolysis liquid penetrate nanofiltration membrane, and polypeptide is cut by nanofiltration membrane
It stays；
The dilute HCl of nanofiltration concentrate is adjusted pH to 6.5 or so, bromelain is added, enzyme concentration is by second of enzymatic hydrolysis
4000U/g substrate digests 60min under the conditions of 50 DEG C, it adds NaOH and adjusts pH to neutrality, product at reduced pressure is concentrated, is spraying
After drying, active peptides are obtained.
The preparation of 2 active peptides of embodiment
Pre-treatment: after sturgeon skin is cut, by scraped cleans such as the subcutaneous fat of fish, connective tissues, then being cut into powder, leaching
It steeps and removes soluble non-collagen in 12wt%NaCl solution, filter out residue and be soaked in isopropanol, ether, distilled water again
According in weight ratio 2:3:7 mixed liquor, fat is removed, is dried after filtering, obtains substrate.
Digest for the first time: substrate be configured to the suspension of 0.3wt% in deionized water, then with dilute HCl adjusting pH to
6.0 or so, bromelain is added, enzyme concentration is 2000U/g substrate, digests 85min under the conditions of 50 DEG C, is then gone out for 100 DEG C
Enzyme 10min adds NaOH and adjusts pH to neutrality, obtains first time enzymolysis product；
Large aperture hyperfiltration treatment: molecular cut off is used to be filtered for the ultrafiltration membrane of 200,000 Da first time enzymolysis product, enzyme
It solves product and penetrates ultrafiltration membrane, the lower albumen of activity and big molecular impurity are retained by ultrafiltration membrane, collect penetrating fluid；
Nanofiltration membrane concentration: ZnCl will be added in ultrafiltration membrane penetrating fluid2, make Zn2+Concentration is divided in 0.2mol/L, then using retention
The nanofiltration membrane of son amount 600Da is filtered concentration, and the polysaccharide, NaCl in enzymolysis liquid penetrate nanofiltration membrane, and polypeptide is cut by nanofiltration membrane
It stays；
The dilute HCl of nanofiltration concentrate is adjusted pH to 6.5 or so, bromelain is added, enzyme concentration is by second of enzymatic hydrolysis
4000U/g substrate digests 65min under the conditions of 55 DEG C, it adds NaOH and adjusts pH to neutrality, product at reduced pressure is concentrated, is spraying
After drying, active peptides are obtained.
The preparation of 3 active peptides of embodiment
Pre-treatment: after sturgeon skin is cut, by scraped cleans such as the subcutaneous fat of fish, connective tissues, then being cut into powder, leaching
Steep and remove soluble non-collagen in 8wt%NaCl solution, filter out residue be soaked in again isopropanol, ether, distilled water by
According in weight ratio 2:3:9 mixed liquor, fat is removed, is dried after filtering, obtains substrate.
Digest for the first time: substrate be configured to the suspension of 0.2wt% in deionized water, then with dilute HCl adjusting pH to
6.0 or so, bromelain is added, enzyme concentration is 2000U/g substrate, digests 75min under the conditions of 50 DEG C, is then gone out for 100 DEG C
Enzyme 10min adds NaOH and adjusts pH to neutrality, obtains first time enzymolysis product；
Large aperture hyperfiltration treatment: molecular cut off is used to be filtered for the ultrafiltration membrane of 400,000 Da first time enzymolysis product, enzyme
It solves product and penetrates ultrafiltration membrane, the lower albumen of activity and big molecular impurity are retained by ultrafiltration membrane, collect penetrating fluid；
Nanofiltration membrane concentration: ZnCl will be added in ultrafiltration membrane penetrating fluid2, make Zn2+Concentration is divided in 0.1mol/L, then using retention
The nanofiltration membrane of son amount 1000Da is filtered concentration, and the polysaccharide, NaCl in enzymolysis liquid penetrate nanofiltration membrane, and polypeptide is cut by nanofiltration membrane
It stays；
The dilute HCl of nanofiltration concentrate is adjusted pH to 6.5 or so, bromelain is added, enzyme concentration is by second of enzymatic hydrolysis
4000U/g substrate digests 55min under the conditions of 55 DEG C, it adds NaOH and adjusts pH to neutrality, product at reduced pressure is concentrated, is spraying
After drying, active peptides are obtained.
The preparation of 1 active peptides of reference examples
Difference with embodiment 1 is: not carrying out secondary enzymolysis processing.
Pre-treatment: it after sturgeon skin is cut, by scraped cleans such as the subcutaneous fat of fish, connective tissues, then is cut into broken
End is soaked in and removes soluble non-collagen in 10wt%NaCl solution, filters out residue and be soaked in isopropanol, ether, steaming again
Distilled water removes fat, dries after filtering, obtain substrate according in weight ratio 3:2:8 mixed liquor.
Digest for the first time: substrate be configured to the suspension of 0.2wt% in deionized water, then with dilute HCl adjusting pH to
6.0 or so, bromelain is added, enzyme concentration is 2000U/g substrate, digests 90min under the conditions of 55 DEG C, is then gone out for 100 DEG C
Enzyme 10min adds NaOH and adjusts pH to neutrality, obtains first time enzymolysis product；
Large aperture hyperfiltration treatment: molecular cut off is used to be filtered for the ultrafiltration membrane of 300,000 Da first time enzymolysis product, enzyme
It solves product and penetrates ultrafiltration membrane, the lower albumen of activity and big molecular impurity are retained by ultrafiltration membrane, collect penetrating fluid；
Nanofiltration membrane concentration: ZnCl will be added in ultrafiltration membrane penetrating fluid2, make Zn2+Concentration is divided in 0.1mol/L, then using retention
The nanofiltration membrane of son amount 800Da is filtered concentration, and the polysaccharide, NaCl in enzymolysis liquid penetrate nanofiltration membrane, and polypeptide is cut by nanofiltration membrane
It stays, nanofiltration concentrate is concentrated under reduced pressure, after spray drying, obtains active peptides.
The preparation of 2 active peptides of reference examples
Difference with embodiment 1 is: ZnCl is not added in ultrafiltrated permeation liquid2。
Pre-treatment: it after sturgeon skin is cut, by scraped cleans such as the subcutaneous fat of fish, connective tissues, then is cut into broken
End is soaked in and removes soluble non-collagen in 10wt%NaCl solution, filters out residue and be soaked in isopropanol, ether, steaming again
Distilled water removes fat, dries after filtering, obtain substrate according in weight ratio 3:2:8 mixed liquor.
Digest for the first time: substrate be configured to the suspension of 0.2wt% in deionized water, then with dilute HCl adjusting pH to
6.0 or so, bromelain is added, enzyme concentration is 2000U/g substrate, digests 90min under the conditions of 55 DEG C, is then gone out for 100 DEG C
Enzyme 10min adds NaOH and adjusts pH to neutrality, obtains first time enzymolysis product；
Large aperture hyperfiltration treatment: molecular cut off is used to be filtered for the ultrafiltration membrane of 300,000 Da first time enzymolysis product, enzyme
It solves product and penetrates ultrafiltration membrane, the lower albumen of activity and big molecular impurity are retained by ultrafiltration membrane, collect penetrating fluid；
Nanofiltration membrane concentration: ultrafiltration membrane penetrating fluid is filtered concentration using the nanofiltration membrane of molecular cut off 800Da, is digested
Polysaccharide, NaCl in liquid penetrate nanofiltration membrane, and polypeptide is retained by nanofiltration membrane；
The dilute HCl of nanofiltration concentrate is adjusted pH to 6.5 or so, bromelain is added, enzyme concentration is by second of enzymatic hydrolysis
4000U/g substrate digests 60min under the conditions of 50 DEG C, it adds NaOH and adjusts pH to neutrality, product at reduced pressure is concentrated, is spraying
After drying, active peptides are obtained.
The preparation of 4 facial mask of embodiment
Prepare raw material: sturgeon fishskin polypeptide 3.875, the Pomegranate Peel Extract being prepared in embodiment 1 according to weight ratio
0.2, guar gum 2.2, hexadecanyl phosphate sylvite 3.2, glycerol 4.5, methyl p-hydroxybenzoate 0.02, essence 0.005, go
Ionized water 86.
A part of deionized water of sturgeon fishskin polypeptide is dispersed, hexadecanyl phosphate sylvite and glycerol, heating is added
To 40-50 DEG C, stir evenly；Guar gum and another part deionized water are continuously added, whitening composition is added, is warming up to 55-
It 60 DEG C, stirs evenly；Methyl p-hydroxybenzoate and essence are added in the solution obtained in stepb, stirs evenly, obtains face
Film liquid；By facial mask liquid be coated on matrix on to get.
Ultra-violet absorption spectrum measurement
Sturgeon polypeptide obtained in embodiment 1 is configured to 0.5 mg/mL aqueous solution, takes 4.0 mL, utilizes UV-Vis spectrophotometry
Photometer is scanned sample within the scope of 200-400 nm.As a result as shown in Figure 1.It can be seen from the figure that SPHs exists
Have strong absworption peak at 220 nm, this is the characteristic absorption peak of peptide bond in proteins and peptides, also have near 270 nm one compared with
Weak absorption peak is the characteristic absorption peak of aromatic amino acid.
The measurement of content of peptides
Sturgeon skin content of peptides is measured using biuret method.Two molecule urea (NH under high temperature3CONH3) reaction sloughs a molecules of ammonia and obtain
To biuret.Polypeptide containing peptide bond can be with the Cu in biuret reagent2+Reaction generates violet complex, and the substance is in 550nm
There is strong absworption peak at place, can be detected with spectrophotometer.This reaction is only related with content of peptides, and product shade contains with polypeptide
It measures directly proportional.
The preparation of biuret reagent: 1.5gCuSO is weighed4•5H2O and 6.0gC4O6H4KNa is dissolved in 500mL deionized water,
It moves into 1L volumetric flask after completely dissolution, the NaOH of 300mL10% is added, constant volume moves into stand-by in brown bottle.By 1.0mL10%
TCA is added in 1.0mL polypeptide solution, and 10min is stood after shaking up, is centrifuged under 4000r/min, 1.0mL supernatant and 4.0mL are taken
Biuret reagent mixing, 50 DEG C of water-bath 10min measure the absorbance value at 550nm.It is drawn with 0 ~ 100mg/mL bovine serum albumin(BSA)
Standard curve processed does reference with pure water.Reference standard curve calculates the content of peptides of sample solution.
The content of peptides that above embodiments and reference examples are prepared is as follows:
As can be seen from the table, the polypeptide content with higher that the present invention is prepared.
Relative molecular mass distribution
Sturgeon skin polypeptide relative molecular weight distribution is measured using efficient liquid phase exclusion chromatography, test condition is as follows: Waters600
High performance liquid chromatograph, 2487 detectors, 220nm wavelength, 30 DEG C of column temperature, flow velocity 0.5mL/min, chromatographic column TSKgel 2000
SWXL, mobile phase are trifluoroethanol: acetonitrile: water=1:450:550.
Wherein the polypeptide molecular weight of embodiment 1, reference examples 1 and reference examples 2 is distributed respectively as shown in figs 2-4, from figure
It can be seen that their range of molecular weight distributions is successively:
As can be seen that the polypeptide molecular weight obtained after secondary enzymolysis in embodiment 1 is smaller, and due to not in reference examples 1
Zn is added in ultrafiltrated permeation liquid2+The activity for resulting in protease in secondary enzymolysis is not high, and range of molecular weight distributions, which is greater than, to be implemented
Example 1, to obtain polypeptide molecular weight larger in the reference examples 2 without carrying out secondary enzymolysis.
The amino acid of sturgeon fishskin polypeptide forms
Sturgeon fishskin polypeptide obtained in embodiment 1 is formed after hydrochloric acid hydrolyzes using its amino acid of high effective liquid chromatography for measuring
And calculate the hydroxyl rate of proline.Determination condition are as follows: Sopium Amino Acid Analysis chromatographic column, linear gradient are washed
De-, A phase is 0.2mol/L sodium citrate aqueous solution (pH3.00), and B phase is 0.2mol/L boric acid sodium water solution (pH9.80), elution
Flow velocity is 0.4mL/min, and column temperature is 65 DEG C.Amino acid composition is as follows:
The measurement of DPPH free radical (DPPH) clearance rate
DPPH is a kind of high chemically active free radical with single electron, and characteristic absorption peak 517nm, alcoholic solution is very
Stablize dark purple.With the single electron of DPPH match reaction can occur for antioxidant, reduce its absorption at 517nm, face
Discoloration is shallow.The sample to be tested of 2.0mL various concentration is drawn, is added 2.0mLDPPH solution (solvent is dehydrated alcohol), after mixing
It is protected from light 1h at 25 DEG C, its absorbance A i is surveyed at 517nm；Control group is 2.0mL ethanol solution and 2.0mL sample, surveys it
Absorbance is Aj；Blank group is 2.0mL ethanol solution and 2.0mLDPPH solution, and surveying its absorbance is A0.
DPPH clearance rate=(1- (Ai-Aj)/A0)×100%
The DPPH free radical scavenging activity for the sturgeon polypeptide that above embodiments and reference examples are prepared is as follows:
As can be seen from the above table, the sturgeon fish-skin that the present invention is prepared has preferably to DPPH free radical scavenging activity, right
As usual due to using Zn to bromelain not during secondary enzymolysis in 22+Auxiliary, it is bad to result in hydrolysis result, makes more
Peptide activity reduces, and only with primary enzymolysis in reference examples 2, so that its activity is bad.
The measurement of hydroxyl radical free radical (OH) clearance rate
Using Fe2+/H2O2Method measures hydroxyl radical free radical clearance rate.Phen-Fe2+There is strong absorption at 536nm, OH can be incited somebody to action
Phen-Fe2+It is oxidized into Phen-Fe3+, weaken the absorption at 536nm.Antioxidant has removing and neutralizes OH
Effect, can inhibit this oxidation process.Sequentially add 0.5mL1.0mmol/L Phen ethanol solution, 0.5mL deionized water,
The FeSO of the PBS and 0.5mL0.75mmol/L of 1.0mL 0.15mol/L pH7.44, it is eventually adding 0.5mL0.01% (v/v)
H2O2, it is put into constant temperature 60min in 37 DEG C of water-baths, surveys its absorbance A in 536nm after reaction sufficientlyi；With the deionized water of 0.5mL
Instead of H2O2Aforesaid operations are repeated, absorbance is denoted as A0；Deionized water repetitive operation is replaced with the sample of 0.5mL, absorbance is denoted as
Aj。
OH clearance rate=(Aj-Ai)/(Ao-Ai)×100%
The hydroxyl radical free radical clearance rate for the sturgeon polypeptide that above embodiments and reference examples are prepared is as follows:
As can be seen from the above table, the sturgeon fish-skin that the present invention is prepared has preferably to hydroxyl radical free radical clearance rate, right
As usual due to using Zn to bromelain not during secondary enzymolysis in 22+Auxiliary, it is bad to result in hydrolysis result, makes more
Peptide activity reduces, and only with primary enzymolysis in reference examples 2, so that its activity is bad.
Nanofiltration membrane is to Na+And Zn2+Ion rejection rate
Using the Na in Atomic Emission Spectrometer AES measurement nanofiltration membrane charging and nanofiltration penetrating fluid+And Zn2+Ion concentration, using such as
Lower formula calculates nanofiltration membrane to the rejection of ion:
R=( Cf -Cp)/Cf×100%
In formula, CfIt is nanofiltration membrane charging intermediate ion concentration, ppm；
CpIt is nanofiltration membrane penetrating fluid intermediate ion concentration, ppm；
R is rejection, %.
Nanofiltration process is as follows to the rejection of ion in the above various embodiments and reference examples:
As can be seen from the table, it uses in the present invention to Zn is added in ultrafiltrated permeation liquid2+Ion is effectively improved to nanofiltration
Film is to Na+Transmitance.
The skin irritation test of facial mask
Using new zealand white rabbit, before carrying out acute skin irritation test, rabbit backbone diamond wool is cut, unhairing range
Left and right Ge Yue 6cm2, formally when experiment, the facial mask being prepared in embodiment 4 is cut to 2.5 × 2.5 cm2Square cover
In on the skin of left side, then covered with two layers of gauze and one layer of glassine paper, then fixed with nonirritant adhesive plaster and bandage, outside
Time 4h is applied, right side skin after the test, cleans skin removed residue with warm water, respectively at 1,24,48h as control
The reaction for observing tested skin carries out skin wound repair scoring using " cosmetics health specification ".It is as follows as the result is shown:
As can be seen from the above table, facial mask of the invention is non-stimulated to skin.
Claims (7)
1. a kind of active peptides facial mask, including matrix and the facial mask liquid being applied in matrix, which is characterized in that the facial mask liquid
In include following component in percentage by weight: sturgeon fishskin polypeptide 2-5%, whitening composition 0.1-0.3%, thickener 2-
4%, emulsifier 3-3.5%, soluble small molecular 4-6%, preservative 0.02-0.04%, essence 0.005-0.01%, deionized water 85-
88%。
2. active peptides facial mask according to claim 1, which is characterized in that the preparation method of the sturgeon fishskin polypeptide
Include the following steps:
Step 1, it pre-treatment: after sturgeon skin is crushed, is soaked in NaCl solution and removes non-collagen, then residue is soaked in
By isopropanol, ether, distilled water mixing and solution in, removal fat, filter out solids；
Step 2, it digests for the first time: solids obtained in step 1 is dispersed in water, HCl is added and adjusts pH to 5.5-6.5,
Then it is digested using bromelain, after enzyme deactivation, adjusting pH to neutrality, obtains first time enzymolysis product；
Step 3, first time enzymolysis product large aperture hyperfiltration treatment: is filtered removal impurity with ultrafiltration membrane；
Step 4, nanofiltration membrane is concentrated: will obtain that ZnCl is added in ultrafiltrated permeation liquid in step 32, reuse molecular cut off 800Da
Nanofiltration membrane be filtered concentration；
Step 5, it digests for second: HCl is added in nanofiltration membrane concentrate obtained in step 4 and adjusts pH to 5.5-6.5, is then made
It is digested with bromelain, after enzyme deactivation, adjusting pH to neutrality, obtains second of enzymolysis product；Successively pass through again
It is concentrated under reduced pressure, spray drying, obtains sturgeon fishskin polypeptide.
3. active peptides facial mask according to claim 1, which is characterized in that the concentration of NaCl solution is 5- in step 1
15%；Isopropanol, ether, distilled water weight ratio be 2-4:1-3:6-10；The concentration of solids in water is 0.1- in step 2
0.5wt%, enzyme concentration are 2000U/g substrates, and hydrolysis temperature is 50-60 DEG C, and enzymolysis time is 60-120min；Ultrafiltration in step 3
The molecular cut off of film is 200,000-40 ten thousand Da；Zn in step 4 in ultrafiltrated permeation liquid2+Concentration is in 0.1-0.5mol/L, nanofiltration
The molecular cut off of film is 500-1000Da；Enzyme concentration is 4000U/g substrate in step 5, and hydrolysis temperature is 45-55 DEG C, enzymatic hydrolysis
Time is 50-70min.
4. active peptides facial mask according to claim 1, which is characterized in that whitening composition is selected from Pomegranate Peel Extract.
5. active peptides facial mask according to claim 1, which is characterized in that thickener is selected from pectin, Sodium Hyaluronate, melon
The mixing of one or more of your glue, xanthan gum, acrylate, carboxymethyl cellulose, cellulose gum or ethyl cellulose.
6. active peptides facial mask according to claim 1, which is characterized in that emulsifier is selected from hexadecanyl phosphate potassium
Salt, stearoylketene base sodium glutamate, one of stearoylketene base-sodium cocoyl glutamate or a variety of；Soluble small molecular is selected from
Glycerol, 1,3-BDO, one of propylene glycol or a variety of；Preservative is methyl p-hydroxybenzoate, P-hydroxybenzoic acid second
One or more of ester, propylparaben, benzoic acid and Phenoxyethanol；The matrix is non-woven fabrics.
7. the preparation method of active peptides facial mask described in claim 1, which comprises the steps of:
Step a disperses a part of deionized water of sturgeon fishskin polypeptide, and emulsifier and soluble small molecular is added, is warming up to
It 40-50 DEG C, stirs evenly；
Step b continuously adds thickener and another part deionized water, adds whitening composition, is warming up to 55-60 DEG C, stirring
Uniformly；
Preservative and essence are added in the solution obtained in stepb, stirs evenly, obtains facial mask liquid by step c；
Step d, by facial mask liquid be coated on matrix on to get.
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| CN111329770A (en) * | 2018-12-18 | 2020-06-26 | 抚顺金达莱生物科技有限公司 | Deep-sea volcano sulfur paste mask cream and preparation method thereof |
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| CN112458139B (en) * | 2020-12-07 | 2022-11-29 | 武汉纺织大学 | Nano active peptide for postpartum repair and production process thereof |
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| KR101582660B1 (en) * | 2014-07-14 | 2016-01-08 | 한국원자력연구원 | Hydrogel Mask Pack Production Containing Collagen Using Radiation |
| CN106191189A (en) * | 2016-09-20 | 2016-12-07 | 武汉百思凯瑞纳米科技有限公司 | A kind of preparation method and applications of tuna skin height moisturizing bioactive peptide |
| CN107648168A (en) * | 2017-10-31 | 2018-02-02 | 福建师范大学 | A kind of filefish fish collagen protein facial mask liquid and preparation method thereof |
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| CN105002246A (en) * | 2015-07-03 | 2015-10-28 | 浙江黛君生物医药科技有限公司 | Fish polypeptide with uniform molecular weight distribution, and preparation method thereof |
| JP6608109B2 (en) * | 2016-03-23 | 2019-11-20 | 富士フイルム株式会社 | Method for enhancing the expression of Endo180 in dermal fibroblasts |
| CN106433486B (en) * | 2016-09-30 | 2018-09-07 | 陕西科技大学 | A kind of preparation method of sturgeon fishskin gelatin |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR101582660B1 (en) * | 2014-07-14 | 2016-01-08 | 한국원자력연구원 | Hydrogel Mask Pack Production Containing Collagen Using Radiation |
| CN106191189A (en) * | 2016-09-20 | 2016-12-07 | 武汉百思凯瑞纳米科技有限公司 | A kind of preparation method and applications of tuna skin height moisturizing bioactive peptide |
| CN107648168A (en) * | 2017-10-31 | 2018-02-02 | 福建师范大学 | A kind of filefish fish collagen protein facial mask liquid and preparation method thereof |
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| CN111329770A (en) * | 2018-12-18 | 2020-06-26 | 抚顺金达莱生物科技有限公司 | Deep-sea volcano sulfur paste mask cream and preparation method thereof |
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